摘要
目的：探讨牙髓炎关键介质白细胞介素（IL）-1β对人牙髓成纤维细胞中基质金属蛋白酶（MMP）-9体外表达水平的影响。方法：体外培养人牙髓成纤维细胞，取4-8代细胞接种于6孔板板孔内作为实验样本，实验组细胞样本培养过程中加入IL-18，对照组不加IL-1β，常规培养。采用链霉素抗生素蛋白-过氧化酶免疫组化法检测并比较实验组和对照组细胞MMP-9免疫组化染色阳性率和染色颗粒分布特点；采用明胶酶谱法检测并比较实验组和对照组细胞MMP-9的明胶酶谱和负染酶带的累积光密度值。结果：免疫组化结果显示，经IL-1β刺激后的人牙髓成纤维细胞中MMP-9的表达增强（P〈0．01），染色阳性颗粒主要分布于细胞胞浆中，少量分布于细胞外基质中；明胶酶谱分析显示，实验组MMP-9水平升高，实验组负染酶带累积光密度值在5个检测时间点均高于对照组（P〈0．01）。结论：IL-1β能够促进人牙髓成纤维细胞中MMP-9的合成和分泌，从而导致牙髓中细胞外基质的降解。
Objective： To investigate the effect of interleukin-1β （IL-1β）, the key factor of dental pulp inflammation, on the synthesis and secretion of matrix metalloproteinases-9 （MMP-9） of human pulp cells in vitro. Methods： Human pulp cells were separated and cultured in vitro. The 4th-8th passage cells were incubated in 6-well plates and divided into two groups. Cells in experimental group were treated with IL-1β, while cells in control group were treated without IL-1β. The streptavidin peroxdase conjugated method was applied to detect immunohistochemical positive stained rate of cells and the distribution of positive stained particles. The gelatin zymography method was applied to examine the production of MMPs in course of culture by the number of integrated optical density. Results： Compared with control, the immunohistochemical re- sult revealed that the expression of MMP-9 was higher in human pulp cells treated with IL-1β（P 〈 0.01）. The immunohisto- chemical positive stained particles were found mainly in the cytoplasm, while a few of them were found in the extracellular ma- trix. Results of gelatin zymography showed that the level of MMP-9 was increased in experimental group than that of control group. Values of integrated optical density were higher in five time points of experimental group than those of control group （P 〈 0.01）. Conclusion： IL-1β has the potential to stimulate the synthesis and secretion of MMP-9 in human pulp cells and thus results in the extracellular matrix degradation of dental pulp.
出处
《天津医药》
CAS
北大核心
2012年第2期142-144,I0004,共4页
Tianjin Medical Journal
